Comparison of Bacillus Thuringiensis with Other Bacillus Species Based on Enzymoserological Examinations of their Proteolytic Enzymes

A selective and differential plating medium, R & F anthracis chromogenic agar (ACA), has been developed for isolating and identifying presumptive colonies of Bacillus anthracis. ACA contains the chromogenic substrate 5-bromo-4-chloro-3-indoxyl-choline phosphate that upon hydrolysis yields teal (blue green) colonies indicating the presence of phosphatidylcholinespecific phospholipase C (PC-PLC) activity. Among seven Bacillus species tested on ACA, only members of the Bacillus cereus group (B. anthracis, B. cereus, and B. thuringiensis) produced teal colonies (PC-PLC positive) having cream rings. Examination of colony morphology in 18 pure culture strains of B. anthracis (15 ATCC strains plus AMES-1-RIID, ANR-1, and AMED-RIID), with one exception, required 48 h at 35 to 37°C for significant color production, whereas only 24 h was required for B. cereus and B. thuringiensis. This differential rate of PC-PLC synthesis in B. anthracis (due to the truncated plcR gene and PlcR regulator in B. anthracis) allowed for the rapid differentiation on ACA of presumptive colonies of B. anthracis from B. cereus and B. thuringiensis in both pure and mixed cultures. Effective recovery of B. anthracis from a variety of matrices having both high (soil and sewage) and low microbial backgrounds (cloth, paper, and blood) spiked with B. anthracis ANR-1 spores suggests the probable utility of ACA plating for B. anthracis recovery in a diversity of applications.

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Di[(R)-2-ethylhexyl] Phthalate, a Bioactive Metabolite First Isolated from Three Different Bacillus Species, and its Synthesis

Letters in Organic Chemistry ◽

10.2174/1570178616666190718125617 ◽

2020 ◽

Vol 17 (2) ◽

pp. 90-95 ◽

Cited By ~ 1

Author(s):

Miriam Castro ◽

Jacqueline Jiménez ◽

Aurelio Ortiz ◽

Estibaliz Sansinenea

Keyword(s):

Biological Activity ◽

Bacillus Thuringiensis ◽

Polyvinyl Chloride ◽

Phthalate Ester ◽

Bacillus Species ◽

Enantiomerically Pure ◽

Bioactive Metabolite ◽

Bacteria And Fungi ◽

First Time ◽

Ethylhexyl Phthalate

: Di(2-ethylhexyl) phthalate (DEHP) is the most common phthalate ester, which has been used as a plasticizer for the production of numerous polymers, particularly polyvinyl chloride (PVC). Many other groups have synthesized meso-DEHP indicating interest in this molecule, but we are the first to synthesize enantiomerically pure di[(R)-2-ethylhexyl] phthalate. We report herein, for the first time, the isolation - from the cultures of Bacillus thuringiensis, B. subtilis, and B. velezensis strains - of di[(R)-2-ethylhexyl] phthalate, enantiomerically pure and in good yields: its biological activity against bacteria and fungi was probed and for the first time its synthesis was done.

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Toxicity of Bacillus species to larvae of Lutzomyia longipalpis (L. & N.) (Diptera: Psychodidae: Phlebotominae)

Anais da Sociedade Entomológica do Brasil ◽

10.1590/s0301-80592000000300025 ◽

2000 ◽

Vol 29 (3) ◽

pp. 609-614 ◽

Cited By ~ 4

Author(s):

Eduardo D. Wermelinger ◽

José C. Zanuncio ◽

Elizabeth F. Rangel ◽

Paulo R. Cecon ◽

Leon Rabinovitch

Keyword(s):

Bacillus Thuringiensis ◽

Visceral Leishmaniasis ◽

Bacillus Sphaericus ◽

Anticarsia Gemmatalis ◽

Bacillus Species ◽

Lutzomyia Longipalpis ◽

Larvicidal Effect ◽

Lepidoptera Noctuidae ◽

Third Instar Larvae

A study was conducted to compare the susceptibility of third instar larvae of Lutzomyia longipalpis (L. & N.) (Diptera: Psychodidae: Phlebotominae), the vector of the American visceral leishmaniasis to two strains of Bacillus thuringiensis serovar israelensis, one strain of Bacillus sphaericus (all pathogenic to Diptera Culicidae) and a strain of B. thuringiensis ser. morrisoni (pathogenic to larvae of Anticarsia gemmatalis (Hübner) (Lepidoptera: Noctuidae)). Larvae of L. longipalpis showed similar susceptibility to the two strains of B. thruringiensis ser. israelensis, while B. sphaericus and B. thuringiensis ser. morrisoni showed low and no larvicidal effect to this vector, respectively.

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Complete Genome Sequence of the Mosquitocidal Bacterium Bacillus sphaericus C3-41 and Comparison with Those of Closely Related Bacillus Species

Journal of Bacteriology ◽

10.1128/jb.01652-07 ◽

2008 ◽

Vol 190 (8) ◽

pp. 2892-2902 ◽

Cited By ~ 61

Author(s):

Xiaomin Hu ◽

Wei Fan ◽

Bei Han ◽

Haizhou Liu ◽

Dasheng Zheng ◽

...

Keyword(s):

Genome Sequence ◽

Complete Genome ◽

Mosquito Control ◽

Proteolytic Enzymes ◽

Biological Control Agent ◽

Sugar Transport ◽

Bacillus Sphaericus ◽

Transport Systems ◽

Bacillus Species ◽

Great Success

ABSTRACT Bacillus sphaericus strain C3-41 is an aerobic, mesophilic, spore-forming bacterium that has been used with great success in mosquito control programs worldwide. Genome sequencing revealed that the complete genome of this entomopathogenic bacterium is composed of a chromosomal replicon of 4,639,821 bp and a plasmid replicon of 177,642 bp, containing 4,786 and 186 potential protein-coding sequences, respectively. Comparison of the genome with other published sequences indicated that the B. sphaericus C3-41 chromosome is most similar to that of Bacillus sp. strain NRRL B-14905, a marine species that, like B. sphaericus, is unable to metabolize polysaccharides. The lack of key enzymes and sugar transport systems in the two bacteria appears to be the main reason for this inability, and the abundance of proteolytic enzymes and transport systems may endow these bacteria with exclusive metabolic pathways for a wide variety of organic compounds and amino acids. The genes shared between B. sphaericus C3-41 and Bacillus sp. strain NRRL B-14905, including mobile genetic elements, membrane-associated proteins, and transport systems, demonstrated that these two species are a biologically and phylogenetically divergent group. Knowledge of the genome sequence of B. sphaericus C3-41 thus increases our understanding of the bacilli and may also offer prospects for future genetic improvement of this important biological control agent.

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Isolation of Tn916-like conjugal elements from swine lot effluent

Canadian Journal of Microbiology ◽

10.1139/w00-029 ◽

2000 ◽

Vol 46 (6) ◽

pp. 542-549 ◽

Cited By ~ 12

Author(s):

Bradley J Haack ◽

Robert E Andrews Jr.

Keyword(s):

Bacillus Thuringiensis ◽

Enterococcus Faecalis ◽

Tetracycline Resistance ◽

Bacillus Species ◽

Pcr Analysis ◽

Soil Environment ◽

Swine Waste ◽

Chain Reaction ◽

Plasmid Pc194 ◽

Polymerase Chain

Isolates of Enterococcus faecalis obtained from a swine farrowing house outflow were examined for genetic elements similar to Tn916. Of the enterococci isolated, 71% were resistant to tetracycline. Among the tetracycline-resistant enterococci isolated from the outflow samples, approximately 34% were able to transfer the tetracycline resistance phenotype to Bacillus thuringiensis in cross-genus matings. The frequencies of transfer for 10 random isolates were comparable to those for transfer of Tn916 from E. faecalis to B. thuringiensis. In addition, these elements were shown to mobilize plasmid pC194 between Bacillus species, as did Tn916. Southern blot and polymerase chain reaction (PCR) analysis showed these elements share extensive structural homology with Tn916. The selected conjugal elements were capable of transfer to a Bacillus recipient in a soil environment. When the swine waste was introduced into the soil, the tetracycline resistant fecal enterococci levels rose from essentially undetectable levels to approximately 4 × 104and remained at this level for 4 weeks. After six months, including one winter, levels had decreased to 5 × 103.Key words: Enterococcus faecalis, Tn916, swine waste, genetic exchange, Bacillus thuringiensis, PCR.

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Sporulation-dependent phagelike particles in inclusion-forming Bacillus species

Canadian Journal of Microbiology ◽

10.1139/m86-072 ◽

1986 ◽

Vol 32 (5) ◽

pp. 373-381 ◽

Cited By ~ 2

Author(s):

Anthony Tam ◽

Philip Fitz-James

Keyword(s):

Electron Microscopy ◽

Bacillus Thuringiensis ◽

Capsid Protein ◽

Late Stage ◽

Immune Reaction ◽

Major Capsid Protein ◽

Stage Ii ◽

Bacillus Species ◽

Mosquito Larvae ◽

Molecular Weights

Phagelike particles, which have been observed in sporulated cells of Bacillus medusa, were found to appear at about late stage II in all supspecies of Bacillus thuringiensis toxic to mosquito larvae. All phagelike particles were similar in size and shape and when purified possessed capsid proteins of similar molecular weights and immune reaction. The synthesis of the major capsid protein occurred at the same time as complete phagelike particles were detected by electron microscopy. The production of phagelike particles was not essential for sporulation.

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Activation and fragmentation ofBacillus thuringiensisδ-endotoxin by high concentrations of proteolytic enzymes

Canadian Journal of Microbiology ◽

10.1139/w99-086 ◽

1999 ◽

Vol 45 (10) ◽

pp. 816-825 ◽

Cited By ~ 6

Author(s):

Anthony SD Pang ◽

J Lawrence Gringorten ◽

Cheng Bai

Keyword(s):

Bacillus Thuringiensis ◽

Insecticidal Activity ◽

Proteolytic Enzymes ◽

Choristoneura Fumiferana ◽

Fragmentation Pattern ◽

Sequencing Data ◽

High Concentration ◽

Sds Page ◽

High Concentrations ◽

Insect Gut

Commercial enzymes and insect gut juice at various concentrations were used to digest Bacillus thuringiensis subsp. sotto Cry1Aa protoxin and examine the fragmentation pattern and effect on insecticidal activity. Trypsin at both high (5 mg/mL) and low (0.05 mg/mL) concentrations converted protoxin to toxin with no difference in insecticidal activity against Bombyx mori larvae. In both cases, the toxin protein had an apparent Mrof 58.4 kDa (SDS-PAGE). Active toxin of identical Mrwas also produced with low concentrations of Pronase and subtilisin, but at high concentration, it was degraded into two protease-resistant fragments of apparent Mr31.8 and 29.6 kDa, and exhibited no insecticidal activity. Sequencing data established the primary cleavage site to be in domain II, the receptor-binding region of the toxin, in an exposed loop between two beta-sheet strands. Fragmentation was not observed, however, when the digests were analyzed by native protein techniques, but rather the toxin molecule appeared to be intact. The amount of activated toxin produced by Choristoneura fumiferana gut juice was markedly reduced when the gut-juice concentration was increased from 1 to 50% and correlated with a loss in insecticidal activity. However, no lower Mrprotease-resistant fragments were evident in the SDS-PAGE of these digests.Key words: Bacillus thuringiensis endotoxin, Lepidoptera, proteolytic enzymes, insect gut juice, activation, digestion.

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